Cell counting kit 8 cck 8

Cell viability was analyzed using the Cell Counting Kit-8 (CCK-8; Servicebio) in accordance with the CCK-8 assay protocols. Cells were seeded into 96-well plates at a density of 2 × 10³ cells/well. When cultured to 0, 24, 48, 72, and 96 h, CCK-8 (10 µL/well) was added to the cells and incubated at 37 °C for 1 h. We measured the absorbance of each well at 450 nm using a microplate reader (Bio-Rad Laboratories, Inc.).

DOX and Cyanine 5.5 (Cy5.5, C41H44N2O14S4) were purchased from Yuanye (Shanghai, China). Fluorescein isothiocyanate/propidium iodide (PI)/calcein AM, 3, 3′-dioctadecyloxacarbocyanine perchlorate (DiO), BCA protein assay kits, and RIPA lysis buffer were purchased from Beyotime Biotechnology (Shanghai, China). Phosphate-buffered saline (PBS), penicillin-streptomycin, fetal bovine serum, and high-glucose Dulbecco’s modified Eagle’s medium were purchased from Thermo Fisher Scientific (Waltham, MA, USA). 4’, 6-diamidino-2-phenylindole (DAPI), phalloidin, phenylmethanesulfonyl fluoride, and Cell Counting kit-8 (CCK-8) were purchased from Service Bio (Wuhan, China). Caspase 9 antibody, BAX antibody, Ki67 antibody, CD3 antibody, CD4 antibody, and β actin antibody were purchased from Proteintech Company (Wuhan, China). Gp100 was purchased from Affinity Biosciences (Santa Clara, CA, USA). B16 cell lines were donated by the National Engineering Laboratory for AIDS Vaccines at Jilin University (24 (link)), and the use of these cell lines received approval from the institutional review board for research ethics at the China-Japan Union Hospital of Jilin University.

Cell viability was assessed using the Cell Counting kit-8 (CCK-8) (Servicebio, G4103, China). Briefly, 5000 HeLa cells per well were seeded into 96well plates. The cells were treated with different chemicals and incubated for different times to evaluate the toxicity of the chemicals. CCK-8 reagent was added to the wells and incubated at 37°C for 2 h. Absorbance was measured at 450 nm using a microplate reader (BioTek 800TS, USA).